Multiplex ligation-dependent probe amplification for the detection of chromosomal gains and losses in formalin-fixed tissue.
until further notice
SourceDiagnostic Molecular Pathology, 14, 1, (2005), pp. 9-16
Article / Letter to editor
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Epidemiology, Biostatistics & HTA
Diagnostic Molecular Pathology
SubjectDCN 2: Functional Neurogenomics; DCN 3: Neuroinformatics; EBP 1: Determinants in Health and Disease; ONCOL 3: Translational research; UMCN 1.2: Molecular diagnosis, prognosis and monitoring; UMCN 1.5: Interventional oncology
Molecular analysis on formalin-fixed paraffin-embedded tissue is of increasing importance in diagnostic histopathology and tumor research. Multiplex ligation-dependent probe amplification (MLPA) is a technique that can be used for detection of copy number alterations of up to 45 different DNA sequences in one experiment. It can be performed on partially degraded DNA, which makes this technique very suitable for analysis of formalin-fixed lesions. We tested the reliability of MLPA by analyzing DNA isolated from formalin-fixed melanomas that were previously characterized by comparative genomic hybridization (CGH), and additionally the applicability of MLPA was tested by analyzing 29 routinely processed melanocytic lesions. MLPA appears to be a reliable and efficient method to evaluate DNA copy number changes as 86% of the loci tested revealed concordant CGH results. Discordance mainly involved alterations that were detected by MLPA and not by CGH probably due to a combination of lower resolution of CGH and occasionally false positive MLPA results. For application of MLPA in a diagnostic setting, different probes on a specific region of interest should be used to prevent false positive MLPA results. In a research setting as well as in a diagnostic setting, MLPA is a fast technique to screen large numbers of formalin-fixed lesions for DNA gains and losses.
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